Toxicological experiments-based evaluation on mutagenicity and teratogenicity of lanthanum nitrate
-
摘要:
目的 评价硝酸镧的致突变性和致畸性，为稀土元素的安全使用提供科学依据。 方法 根据硝酸镧对ICR小鼠的半数致死剂量（LD50），分别设高、中、低（雄性为875、437.5、218.75 mg/kg·BW；雌性为1 015 、507.5、253.75 mg/kg·BW）3个剂量组，对ICR小鼠（每组 10 只，雌雄各半）进行骨髓细胞微核试验，观察硝酸镧对小鼠骨髓微核细胞率的影响；根据硝酸镧对CHL细胞的半数抑制浓度，分别设高、中、低（不加S9混合液时为1 × 10 – 3、1 × 10 – 4、1 × 10 – 5 mol；加S9混合液时为5 × 10 – 3、2.5 × 10 – 4、1.25 × 10 – 5 mol）3个剂量组，进行体外哺乳动物细胞（接种至25 mL细胞培养瓶中对数分裂期生长良好的 CHL 细胞，1 × 106 个/瓶）染色体畸变试验，观察硝酸镧对哺乳动物细胞染色体畸变率的影响；设硝酸镧5个剂量组（分别为5、1、0.2、0.04、0.008 mg/皿），在S9代谢活化系统存在或不存在的条件下进行鼠伤寒沙门氏菌（TA97、TA98、TA100、TA102、TA1535）回复突变试验，观察硝酸镧对鼠伤寒沙门氏菌回复突变的影响。设硝酸镧3个剂量组（分别为9、36、144 mg/kg·BW），通过对SD孕鼠（每组至少 15 只）的致畸试验，观察硝酸镧对孕鼠生长和着床能力、胎仔体重和身长情况及对胎鼠骨骼畸形率、各脏器畸形率及总内脏畸形率的影响。 结果 小鼠骨髓细胞微核试验结果显示，硝酸镧各剂量组的微核细胞率与阴性对照组比较，差异无统计学意义（P > 0.05）。体外哺乳动物细胞染色体畸变试验结果显示，在不加S9混合液的条件下，硝酸镧在1 × 10 – 5 M时可导致染色体畸变细胞率升高（χ2 = 12.109，P < 0.01），但畸变细胞率 < 5%，可能不具有生物学意义；在加S9的条件下，硝酸镧各剂量组的染色体畸变细胞率与阴性对照组比较，差异无统计学意义（P > 0.05）。鼠伤寒沙门氏菌回复突变试验结果显示，在S9代谢活化系统存在或不存在的条件下，硝酸镧各剂量组的回复突变菌落数与阴性对照组比较均无明显升高（P > 0.05）。大鼠致畸试验结果显示，硝酸镧各剂量组（144、36、9 mg/kg·BW）对孕鼠生长、着床能力均无明显影响；高剂量组（144 mg/kg·BW）胎仔体重、身长均明显低于阴性对照组（P < 0.05）、胎鼠骨骼畸形率较阴性对照组显著升高（χ2 = 9.939，P < 0.05），各剂量组胎鼠各脏器畸形率及总内脏畸形率与阴性对照组比较差异无统计学意义（P > 0.05）。 结论 在本试验条件下，硝酸镧无明显致突变性；硝酸镧最大无致畸作用剂量为36 mg/kg·BW（以镧元素计为15.38 mg/kg·BW）。 Abstract:Objective To evaluate mutagenic and teratogenic effects of lanthanum nitrate for providing evidence to safe use of the rare earth element. Methods The mutagenicity of lanthanum nitrate was evaluated by micronucleus assay on mouse bone marrow cells, in vitro chromosome aberration test, and Salmonella typhimurium strains bacterial reverse mutation test and the teratogenic effect of lanthanum nitrate was assessed with teratogenicity test in rats. The micronucleus assay on mouse bone marrow cells was performed in three groups of ICR mice (10 in each group, half male and half female) administrated with lanthanum nitrate at concentrations (mg/kg·BW) of 875, 437.5, 218.75 for the male mice and 1015, 507.5, 253.75 for the female mice according to the median lethal dose (LD50) of lanthanum nitrate for the mice. The chromosome aberration test of mammalian cells in vitro was conducted on Chinese hamster lung (CHL) cells (with good growth during logarithmic division and inoculated into 25 mL cell culture bottle, 1 × 106 cells per bottle) treated with lanthanum nitrate at concentrations of 1 × 10 – 3, 1 × 10 – 4, 1 × 10 – 5 mol (without S9 mixture) and 5 × 10 – 3, 2.5 × 10 – 4,1.25 × 10 – 5 mol (with S9 mixture) based on the median inhibition concentration (IC50) of lanthanum nitrate on CHL cells. Five lanthanum nitrate dose groups (5, 1, 0.2, 0.04, 0.008 mg/dish, respectively) were set up to carry out Salmonella typhimurium (TA97, TA98, TA100, TA102, TA1535) strains bacterial reverse mutation test in the presence or absence of S9 metabolic activation system. In teratogenicity test on pregnant Sprague-Dawley (SD) rats (at least 15 in each group), three lanthanum nitrate dosages of (9, 36, 144 mg/kg·BW) were determined and the growth and implantation ability and the offspring rats′ weight and length, rate of skeletal malformation, rate of organ malformation and total rate of viscera malformation were oberserved. Results In micronucleus assay, the micronucleus formation rates of bone marrow cells in each lanthanum nitrate dose group were similar with that of vehicle control group (P > 0.05) . In vitro chromosome aberration test, the rate of cells with chromosome aberration in 1 × 10 – 5 M lanthanum nitrate group significantly increased in the absence of S9 compared to that of the control group (χ2 = 12.109, P < 0.01); however, the proportion of cells with chromosome aberration was less than 5%, indicating a biological insignificance; in the presence of S9, there was no significant difference in chromosome aberration the between each lanthanum nitrate dose group and the control group (all P > 0.05). The results of Salmonella typhimurium strains bacterial reverse mutation test showed that there were no significant differences in the mutagenicity among lanthanum nitrate dose groups and the control group in both the presence and absence of S9 mixture. The results of teratogenic test showed that there were no significant differences in the growth and the implantation ability of pregnant rats among all dose groups and the vehicle group; lanthanum nitrate at concentration of 144 mg/kg BW inhibited fetal body weight and length significantly (both P < 0.05) and increased skeletal malformation of fetal rat compared with those of the vehicle group (χ2 = 9.939, P < 0.05). However, there was no significant difference in organ-specific and total visceral malformation among the dose groups (P > 0.05 for all). Conclusions The mutagenic effect of lanthanum nitrate was not observed under the conditions of this study and the study resulted in a no observable adverse effect level (NOAEL) of 36 mg/kg BW for lanthanum nitrate (15.38 mg/kg BW in terms of lanthanum). -
Key words:
- lanthanum nitrate /
- mutagenicity /
- teratogenicity /
- safety evaluation
-
表 1 硝酸镧对小鼠骨髓微核细胞率的影响（$\bar x \pm s$）
Table 1. Micronucleus rate of bone marrow cells by gender and dose in mice treated with lanthanum nitrate at concentrations of 875.0, 437.5, 218.75 mg/kg·BW for male mice and 1 015.0, 507.5, 253.75 mg/kg·BW for female mice
小鼠 组别 小鼠数 受检PCE
总数（个）含微核 PCE数 PCE NCE PCE/NCE 雌性 阴性对照组 5 10 000 3 611 389 1.58 ± 0.12 低剂量组 5 10 000 2 569 431 1.35 ± 0.28 中剂量组 5 10 000 7 570 430 1.34 ± 0.22 高剂量组 5 10 000 4 591 409 1.44 ± 0.07 阳性对照组 5 10 000 190 a 629 371 1.71 ± 0.22 雄性 阴性对照组 5 10 000 2 620 380 1.63 ± 0.12 低剂量组 5 10 000 2 595 405 1.47 ± 0.15 中剂量组 5 10 000 2 572 428 1.36 ± 0.24 高剂量组 5 10 000 3 609 371 1.56 ± 0.10 阳性对照组 5 10 000 190 b 645 355 1.82 ± 0.15 注：与阴性对照组比较，a P < 0.01; 与阴性对照组比较，b P < 0.01。 
下载: 导出CSV
表 2 硝酸镧对CHL细胞染色体畸变的影响
Table 2. Chromosome aberration of CHL cells treated with lanthanum nitrate at different concentrations
S9混合液 组别 观察细胞数 畸变细胞数 染色体畸变细胞率（%） – S9 阴性对照组 300 5 1.7 ± 0.6 硝酸镧组（M） 1 × 10 – 5 300 14 4.7 ± 0.6 a 1 × 10 – 4 300 4 1.3 ± 0.6 1 × 10 – 3 300 3 1.0 ± 0.0 丝裂霉素C组（1.0 μg/ml） 300 165 55.0 ± 6.6 b + S9 阴性对照组 300 5 硝酸镧组（M） 1.25 × 10 – 5 300 5 1.7 ± 1.5 2.50 × 10 – 4 300 3 1.0 ± 0.0 5.00 × 10 – 3 300 3 1.0 ± 0.0 环磷酰胺组（9.5 μg/ml） 300 170 56.7 ± 6.4 c 注：与阴性对照组比较，a P < 0.01；与阴性对照组比较，b P < 0.01；与阴性对照组比较，c P < 0.01。
下载: 导出CSV
表 3 硝酸镧对鼠伤寒沙门氏菌自发回变数的影响
Table 3. Spontaneous regression variables of Salmonella typhimurium cultured with
lanthanum nitrate of different concentrations 组别 TA97 TA98 – S9 + S9 – S9 S9 溶剂对照组
硝酸镧组（mg/皿）102.3 ± 5.1 93.0 ± 4.0 30.7 ± 3.1 30.0 ± 3.5 0.008 100.0 ± 5.6 83.7 ± 4.2 28.7 ± 4.7 27.3 ± 3.8 0.04 99.7 ± 7.5 82.7 ± 5.5 28.3 ± 2.5 25.0 ± 2.6 0.2 92.0 ± 8.5 84.0 ± 7.9 30.0 ± 1.0 29.7 ± 3.8 1.0 103.0 ± 4.4 90.7 ± 7.1 28.0 ± 3.0 29.7 ± 4.5 5.0 96.0 ± 3.5 79.7 ± 6.1 25.3 ± 3.8 23.7 ± 3.1 阳性对照组 1149.3 ± 74.3 898.7 ± 68.0 490.7 ± 18.0 472.0 ± 65.5
下载: 导出CSV
组别 TA100 TA102 TA1535 – S9 + S9 – S9 + S9 – S9 + S9 溶剂对照组
硝酸镧组（mg/皿）123.7 ± 5.7 150.7 ± 6.0 290.7 ± 15.5 302.7 ± 10.1 11.0 ± 1.0 10.7 ± 0.6 0.008 136.0 ± 7.9 147.0 ± 3.6 276.7 ± 18.6 287.0 ± 5.0 10.3 ± 1.5 9.7 ± 1.5 0.04 128.0 ± 5.3 141.3 ± 4.0 276.7 ± 6.7 283.0 ± 11.8 10.3 ± 2.1 10.7 ± 2.5 0.2 138.0 ± 7.5 142.0 ± 3.6 275.0 ± 7.5 284.0 ± 4.6 11.0 ± 1.7 9.7 ± 2.1 1 136.3 ± 4.5 145.0 ± 10.5 264.0 ± 6.0 276.0 ± 9.2 11.0 ± 2.0 1 0.7 ± 0.6 5 114.7 ± 9.5 150.7 ± 12.2 259.7 ± 14.6 275.7 ± 9.5 11.0 ± 2.6 9.0 ± 1.0 阳性对照组 768.0 ± 57.7 832.0 ± 56.0 994.7 ± 72.6 789.3 ± 53.3 420.7 ± 51.1 100.3 ± 3.5
下载: 导出CSV
表 4 硝酸镧对胎鼠发育的影响（$ \overline {\text{X}} $ ± s）
Table 4. Impacts of lanthanum nitrate of different concentrations on development of fetal rats
组别 胎仔数（只） 胎盘重（g） 胎鼠体重（g） 胎鼠身长（cm） 胎鼠尾（cm） 阴性对照组 210 0.54 ± 0.04 3.91 ± 0.18 3.62 ± 0.08 1.33 ± 0.07 硝酸镧组（mg/kg·BW） 9 191 0.52 ± 0.03 3.98 ± 0.28 3.98 ± 0.28 1.35 ± 0.06 36 201 0.53 ± 0.05 3.91 ± 0.21 3.91 ± 0.21 1.34 ± 0.04 144 219 0.52 ± 0.06 3.72 ± 0.23 a 3.53 ± 0.12 b 1.34 ± 0.05 阳性对照组 259 0.56 ± 0.04 3.98 ± 0.21 3.55 ± 0.10 c 1.34 ± 0.06 注：与阴性对照组比较，a P < 0.01；与阴性对照组比较，b P < 0.05；与阴性对照组比较，c P < 0.05。
下载: 导出CSV
表 5 硝酸镧对胎鼠骨骼发育的影响
Table 5. Impacts of lanthanum nitrate of different concentrations on skeletal development of fetal rats
组别 胎仔数 顶骨骨化不全 枕骨骨化不全 肋骨畸形 尾骨畸形 骨骼畸形 数（只） 畸形率（%） 数（只） 畸形率（%） 畸形数（只） 畸形率（%） 畸形数（只） 畸形率（%） 数（只） 畸形率（%） 阴性对照组 112 3 2.7 5 5.4 0 0 0 2.8 9 8 硝酸镧（mg/kg·BW） 9 102 10 9.8 7 8.8 1 1 1 1.5 19 18.6 36 109 9 8.3 7 12.8 1 0.9 1 4.7 23 21.1 144 116 10 8.6 8 12.1 0 0 0 4.4 26 22.4 b 阳性对照组 137 15 a 10.9 11 13.9 4 2.9 4 3.7 35 25. c 注：与阴性对照组比较，a P < 0.05；与阴性对照组比较，b P < 0.05；与阴性对照组比较，c P < 0.01。
下载: 导出CSV
-
[1] 李睿珺, 周雅琳, 刘伟, 等. 稀土元素对动物的生物学效应研究进展[J]. 环境与健康杂志, 2020, 37(4): 1008 – 1014. [2] 鲍理涛. 中国稀土出口现状、问题及建议[J]. 对外经贸, 2019(7): 20 – 22. [3] Paulick H, Machacek E. The global rare earth element exploration boom: an analysis of resources outside of China and discussion of development perspectives[J]. Resources Policy, 2017, 52: 134 – 153. doi: 10.1016/j.resourpol.2017.02.002 [4] 陈祖义, 朱旭东, 王筱霏. 茶叶稀土含量对植物性食品质量安全的影响[J]. 中国食品卫生杂志, 2016, 28(2): 263 – 268. [5] Xie F, Zhang TA, Dreisinger D, et al. A critical review on solvent extraction of rare earths from aqueous solutions[J]. Minerals Engineering, 2014, 56: 10 – 28. doi: 10.1016/j.mineng.2013.10.021 [6] 汤猛猛, 葛建鸿, 曹凯鑫, 等. 硝酸镧对成年雌性小鼠免疫功能及细胞病理学的影响[J]. 环境与职业医学, 2020, 37(5): 447 – 452. [7] Wang XY, Tang MM, Ge JH, et al. Effects of intrauterine and lactational exposure to lanthanum nitrate on BALB/c offspring mice: developmental immunotoxicity and self-recovery[J]. Toxic-ology Letters, 2022, 362: 17 – 25. doi: 10.1016/j.toxlet.2022.01.013 [8] Pagano G, Thomas PJ, Di Nunzio A, et al. Human exposures to rare earth elements: present knowledge and research prospects[J]. Environmental Research, 2019, 171: 493 – 500. doi: 10.1016/j.envres.2019.02.004 [9] Paulick H, Machacek E. The global rare earth element exploration boom: an analysis of resources outside of China and discussion of development perspectives[J]. Resources Policy, 2017, 52: 134 – 153. doi: 10.1016/j.resourpol.2017.02.002 [10] 纪云晶, 崔明珍, 王贻嘉, 等. 农用稀土硝酸盐毒理学评价[J]. 中华劳动卫生职业病杂志, 1987, 5(1): 54 – 56. [11] Zhou L, Li SL, Chen H, et al. DNA damage effect to mixed rare earth Changle crossing placenta barrier on rat embryo[J]. Journal of Rare Earths, 2003, 21(2): 176 – 179. [12] 杨辉, 嵇庆, 张锡然, 等. 氯化钇和氯化镨引起的人淋巴细胞DNA分子损伤的研究[J]. 遗传, 1998, 20(2): 16 – 18. [13] Oral R, Pagano G, Siciliano A, et al. Heavy rare earth elements affect early life stages in Paracentrotus lividus and Arbacia lixula sea urchins[J]. Environmental Research, 2017, 154: 240 – 246. doi: 10.1016/j.envres.2017.01.011 [14] Trifuoggi M, Pagano G, Guida M, et al. Comparative toxicity of seven rare earth elements in sea urchin early life stages[J]. Environmental Science and Pollution Research, 2017, 24(25): 20803 – 20810. doi: 10.1007/s11356-017-9658-1 [15] 杨飞玉, 王丽, 高艳荣, 等. 氯化镧短期重复剂量染毒对ICR小鼠遗传毒性和氧化损伤的影响[J]. 包头医学院学报, 2014, 30(4): 9 – 11. [16] 杨业鹏, 贾凤兰, 蒋建军, 等. 硝酸镧灌胃3个月对大鼠骨髓细胞的致染色体畸变作用[J]. 环境与职业医学, 2002, 19(3): 198. [17] 康陈萍, 刘青云, 肖倩倩, 等. 利用全胚胎和微团培养模型评价硝酸镧的发育毒性[J]. 癌变·畸变·突变, 2020, 32(4): 256 – 263, 268. [18] Fang HQ, Yu Z, Zhi Y, et al. Subchronic oral toxicity evaluation of lanthanum: a 90-day, repeated dose study in rats[J]. Biomedical and Environmental Sciences, 2018, 31(5): 363 – 375. [19] 马新群, 陆丹, 陈志莲, 等. SD大鼠血液学指标和血清生化学指标参考值探讨[J]. 实用预防医学, 2006, 13(6): 1467 – 1468. [20] 刘志宏, 雷志明, 魏雪涛, 等. 硝酸稀土经乳汁对小鼠仔代免疫功能的影响[J]. 中华预防医学杂志, 2002, 36(6): 394 – 397. [21] 戚大煜, 蔡原, 巫生文. 哺乳期母鼠镧暴露对子代脑组织氧化损伤影响[J]. 中国公共卫生, 2009, 25(6): 704 – 705. [22] Huang J, Zhang TL, Xu SJ, et al. Effects of lanthanum on composition, crystal size, and lattice structure of femur bone mineral of wistar rats[J]. Calcified Tissue International, 2006, 78(4): 241 – 247. doi: 10.1007/s00223-005-0294-2 [23] Liu DD, Zhang JC, Wang GF, et al. The dual-effects of LaCl3 on the proliferation, osteogenic differentiation, and mineralization of MC3T3-E1 cells[J]. Biological Trace Element Research, 2012, 150(1 – 3): 433 – 440. [24] Shin SH, Kim HO, Rim KT. Worker Safety in the rare earth elements recycling process from the review of toxicity and issues[J]. Safety and Health at Work, 2019, 10(4): 409 – 419. doi: 10.1016/j.shaw.2019.08.005 [25] 中华人民共和国卫生部. 化学品毒性鉴定技术规范[R]. 北京: 中国劳动社会保障出版社, 2005: 136 – 141. [26] 周蕊, 刘丽萍, 岳兵, 等. 第四次中国总膳食研究 – 膳食稀土元素的摄入量[J]. 现代预防医学, 2014, 41(11): 1975 – 1978. -
点击查看大图
表(6)
计量
- 文章访问数: 23
- HTML全文浏览量: 8
- PDF下载量: 3
- 被引次数: 0